PP204 ENTERAL INFUSION OF GLUCOSE MODULATES HUMAN DUODENAL PROTEOME BUT DOES NOT ALTER TOTAL PROTEIN TURNOVER

102 PP202 THE INTEGRIN/SRC SYSTEM: AN ESSENTIAL COMPONENT IN L-GLUTAMINE-MEDIATED CELLULAR PROTECTION, HSP70 EXPRESSION, AND O-GLYCOSYLATION VIA CELL SWELLING S. Niederlechner1 , C. Hamiel1 , P. Wischmeyer1 . 1 Anesthesiology, University of Colorado, Denver, United States Rationale: It is known that Glutamine (GLN) can induce protective heat shock proteins (HSP) by the O-glycosylation (O-GlcNAc) pathway and that cell swelling and osmosensing pathways can induce cellular protection. We investigated whether this effect may be related to GLN-mediated cell swelling and whether the integrin/Src pathway is part of GLN’s cell protective mechanism. Methods: Intestinal epithelial-6 cells treated for 15min with increasing doses of GLN up to 20mM, with/without the integrin inhibitor GRGDSP (50mM), inactive control peptide GRGESP (50mM) or Src-kinase inhibitor PP-2 (75mM). Cell survival via MTS assay 24 h post-heat shock (HS) (44ºC × 50 min). O-glycosylated protein levels and HSP70 levels determined via Western blot and cell size via fluorescence microscopy after non-lethal HS (43ºC × 50 min). Results: GLN increased cell survival and cell size in a dose dependent manner (p < 0.001 vs. HS CT; n = 4). GRGDSP completely attenuated GLN-mediated cell swelling in all groups (p < 0.001 vs. HS GLN; n = 4). GLN’s protection was completely attenuated by GRGDSP in 2mM GLN groups and was decreased by 88% and 85% in 10mM and 20mM GLN groups (p < 0.001 vs. respective GLN groups; n = 4). PP-2 attenuated GLN’s protection by 86% in the 2mM GLN group, by 77% and 70% in 10mM and 20 mM GLN groups (p < 0.001 vs. respective GLN groups; n = 4). O-GlcNAc modified proteins increased 100% post-HS after 10mM GLN (p < 0.001 vs. HS C; n = 5). GRGDSP attenuated 10mM GLN’s effect on O-GlcNAc protein modification by 84% (p < 0.05 vs. HS 10mM GLN; n = 5). HSP70 increased by 119% in the 10mM GLN group after HS (p < 0.001 vs. HS CT; n = 4). GRGDSP attenuated HSP70 expression in this group by 88% (p < 0.05 vs HS GLN; n = 4). GRGESP had no effect on GLN-mediated cell protective pathways. Conclusion: The integrin/Src pathway is an essential component of GLN’s protective molecular mechanism. Supported: R01 GM078312 Disclosure of Interest: None declared

PP203 A HIGH WHEY-PROTEIN LEUCINE-ENRICHED LOWCALORIC SUPPLEMENT RESULTS IN HIGHER AND FASTER RISE IN SERUM AMINO ACID LEVELS THAN A CASEIN CONTAINING OR HIGH-CALORIC SUPPLEMENT EQUIVALENT Y.C. Luiking1 , R. Memelink1 , Y. Boirie2 , G. Verlaan1 . 1 Centre for Specialized Nutrition, Danone Research, Wageningen, Netherlands; 2 Unit´ e de Nutrition Humaine, UMR INRA/Universit´ e d’Auvergne, Clermont-Ferrand, France Rationale: Compared to younger adults, elderly need higher levels of blood amino acids (AA), especially leucine

Poster presentations and essential AA (EAA), to stimulate muscle protein synthesis. This study aimed to evaluate the acute effect of high-protein oral nutritional supplements varying in protein source and caloric density on serum AA levels in elderly. Methods: 12 healthy elderly (65 70 y, BMI 22 30 kg/m2 ) participated in this randomised, controlled, single blind, cross-over study. On 4 separate occasions with a washout period of at least one week, subjects consumed iso-nitrogenous (21 g protein) supplements containing: whey+leucine, 150 kcal (W150); casein, 150 kcal (C150); whey+leucine, 320 kcal (W320); and casein, 320 kcal (C320). Blood samples were taken immediately before and at 15 to 30-min intervals until 4 h after product intake for serum AA levels. Mixed model ANOVA was used for statistics. Data are means±SEM. Results: Maximum leucine concentration was significantly higher for W150 vs. C150 (521±15 vs. 260±15 mmol/L, p < 0.001), and for W320 vs. C320 (406±15 vs. 228±15 mmol/L, p < 0.001). Maximum leucine concentration was higher for the low-caloric vs. the high-caloric products (p < 0.001 for pooled analyses), with interaction between protein source and caloric density (p < 0.001). Similar effects were observed for the incremental area under the curve (iAUC) and for maximum concentrations of total AA and EAA. Time to reach half the iAUC (t1/2 ) was significantly shorter for W150 vs. C150 (78±5 vs. 101±5 min, p < 0.005) and for W320 vs. C320 (103±5 vs. 87±5 min, p < 0.01). Conclusion: The whey+leucine containing products provide a faster rise in serum AA and result in higher serum levels of total AA, EAA and leucine, further enhanced by a low-caloric density. Therefore, a whey+leucine product seems preferable to provoke muscle protein synthesis in elderly. Disclosure of Interest: None declared

PP204 ENTERAL INFUSION OF GLUCOSE MODULATES HUMAN DUODENAL PROTEOME BUT DOES NOT ALTER TOTAL PROTEIN TURNOVER A. Goichon1 , M. Co¨ effier1 , S. Claeyssens2 , S. Lecleire3 , 4 A.-F. Cailleux , P. Chan5 , N. Donnadieu6 , A. Lavoinne2 , O. Boyer7 , D. Vaudry5 , P. D´ echelotte1 . 1 ADEN EA4311, Institute for Biomedical Research, IFRMP23, Rouen University, 2 Laboratory of Medical Biochemistry, 3 Department of Gastroenterology, 4 Clinical Investigation Center CIC 0204-INSERM, Institute for Biomedical Research, IFRMP 23, Rouen University Hospital, 5 Platform in Proteomics and INSERM U982, IFRMP 23, Rouen University, 6 Department of Pharmacy, Rouen University Hospital, 7 INSERM U905, Institute for Biomedical Research, IFRMP23, Rouen University, Rouen, France Rationale: Glucose (Glc) is a major energy substrate for human enterocytes. Therefore, we aimed to assess the effects of enteral Glc on human duodenal proteome and protein turnover. Methods: The study was approved by the local ethics committee. Twenty healthy volunteers received a 5 h enteral infusion of either saline (NaCl) or glucose (0.67 mmol.kg 1 .h 1 ), and a continuous intravenous infu-

Protein and amino acid metabolism sion of L-[1-13 C]-leucine (12 mmol.kg 1 .h 1 ) before endoscopic duodenal biopsies were taken. Duodenal protein fractional synthesis rate (FSR) was calculated from GC/MS-assessed enrichments in protein and free amino acid pool, and proteolytic activities were evaluated using specific fluorogenic substrates. A 2D-PAGE-based comparative proteomics analysis from biopsies was also performed and differentially expressed proteins were identified by LC-MS/MS. Protein expressions were confirmed by western blotting analysis. Results were compared using MannWhitney or t-test as appropriate, p < 0.05 was considered significant. Results: Duodenal protein FSR [55±5% vs 64±9%/day] and proteolytic activities were not affected by Glc infusion compared to saline, respectively. However, the comparative proteomics analysis indicated that 10 protein spots were differentially (i.e. at least±1.5 fold) and significantly (p < 0.05; t-test) expressed in response to the Glc infusion vs NaCl. The eight identified proteins were mainly implicated in metabolism pathways. For example, a-enolase, aconitase 1 and glutathione S-transferase w-1 were upregulated, while epoxide hydrolase 2 was downregulated. Conclusion: An enteral infusion of Glc does not affect total duodenal protein synthesis or proteolysis in healthy humans, but modulates the duodenal proteome by inducing or inhibiting the expression of enzymes involved mainly in carbohydrate and xenobiotic metabolisms. Disclosure of Interest: None declared

PP205 EFFECTS OF LEUCINE-SUPPLEMENTED WHEY PROTEIN ON GROWTH RATE OF YOUNG RATS P.C. Lollo1 , I.G. Boretti1 , T.M. Batista2 , E.M. Carneiro2 , N.M. Povel1 , J. Amaya-Farfan1 . 1 Food and Nutrition, 2 Institute of Biology, Unicamp, Campinas, Brazil Rationale: Intro/Aim In both animals and humans L-leucine mediates activation of protein synthesis, but the rate of growth as a function of dose is subject to variation, depending on the type of protein. The purpose of this study was to assess growth rate, mTOR activation and liver function in rats fed whey protein (WP) supplemented with leucine at four levels (WP+0, +3, +4.5 and +6% of diet energy adjusted at the expense of carbohydrate). Methods: Twenty four male Wistar rats were divided into four groups (n = 6) and fed one of the following for 30 days, modified AIN 93-G diets containing whey protein instead of casein: WP (whey protein, and WP+4.5 (4.5% L-leucine). Student’s t test was applied to compare the means (significance p < 0.05). Results: Growth peaked at L-leucine 4.5% (128.4% weight increment in 30 days or 2.79%/day), against 97.9% for the control group (WP+0) (p > 0.05). Diet supplementation with L-leucine did not adversely affect liver function (AST, ALT) or body weight. mTOR activation did not differ between groups WP+0 and WP+4.5, but protein level of carcass was greatest at 4.5% supplementation (WP+0 = 41.2%; WP+4.5 = 45.2; p < 0.05). Conclusion: Supplementation with 4.5% L-leucine of diet with whey protein as protein source may permit maximal

103 growth at lower protein intakes, while exhibiting a higher body protein concentration. Disclosure of Interest: None declared

PP206 SINGLE ADMINISTRATION OF HIGH DIETARY GLYCATION PRODUCTS DOES NOT BLUNT ANABOLIC RESPONSE OF SKELETAL MUSCLE IN ADULT RATS H. Murakami1,2 , G. Henry3 , J. L´ eonil3 , M.-J. Galmier4 , J.-M. Chezal4 , C. Giraudet1,2 , A. Masgrau1,2 , S. Warland1,2 , Y. Boirie1,2,5 , C. Guillet1,2 . 1 UMR 1019 Human Nutrition Unit, INRA-Universite d’Auvergne, 2 CRNH Auvergne, Clermont-Ferrand, 3 UMR 1253 Science et Technologie du Lait et de L’Oeuf (STLO), INRA-Agrocampus Ouest, Rennes, 4 UMR 990 Laboratoire de Chimie Analytique et Spectrometrie de Mass, Inserm/Universite d’Auvergne, 5 CHU, Gabriel Montpied Hospital, Clermont-Ferrand, France Rationale: Chronic intake of high dietary glycation products (GP) is associated with some disorders such as insulin resistance in aging and diabetes. We hypothesize that single administration of dietary GP may affect insulin signaling pathway in skeletal muscle. Methods: Semi-liquid solution of diet containing heated casein (high GP) or non-heated casein (low GP) was single administered in 7 months old rats. Plasma insulin was measured by ELISA, and phosphorylation of Akt, p70S6K protein and receptor of GP (RAGE) were measured by Western-blot in soleus (type I) and tibialis (type II) at 0, 1, 2, 4 hours after diet administration (n = 4 or 5 in each time and group). Results: Plasma insulin increased after diet administration, but did not differ between groups. After low GP diet administration, phosphorylation of Akt and p70S6K were significantly increased in tibialis, but not in soleus. RAGE expression was not changed in tibialis and soleus. Phosphorylation of p70S6K and RAGE expression were not different between low and high GP in each time in tibialis and soleus. Phosphorylation of Akt was significantly increased in soleus at 1 hour after the administration of high dietary GP (1.9 fold, p < 0.05). Conclusion: Single administration of high dietary glycation products does not impair insulin and protein translation signaling pathway in type I and type II skeletal muscle in adult rats. Consequently, these data suggest that postprandial anabolic response of skeletal muscle is maintained after single intake of high dietary glycation products. Disclosure of Interest: None declared

PP207 NITROGEN BALANCE AND PROTEIN REQUIRIMENT FOR AH1N1 INFLUENZA PATIENTS E. Perez Cruz1 , N.J. Castillo Garcia1 , E. Gonzalez Aguilar1 . 1 Apoyo Nutricio, Hospital Juarez de Mexico, Mexico DF, Mexico Rationale: The AH1N1 influenza emerged in 2009 as a pandemic with high mortality rate. These patients have an interesting metabolic behavior, where energy demands seem to have a marked increase and a severe catabolism.